Method for extracting protein from kaniwa and protein products thereof

ABSTRACT

Provided herein is a method for making a protein product from kaniwa. The method comprises milling the kaniwa. The milled kaniwa is contacted with a salt solution. A protein is precipitated from the kaniwa. The precipitated protein is dried to form a protein product. Also provided herein is a protein product produced by a method disclosed herein.

CROSS-REFERENCE

The present disclosure claims priority to U.S. Provisional ApplicationSer. No. 61/770,716 filed Feb. 28, 2013, and entitled “Method forExtracting Protein from Kaniwa and Protein Products Thereof,” which isincorporated herein by reference in its entirety.

TECHNICAL FIELD

The present disclosure relates to protein extraction from a seed orgrain, in particular to protein extraction and a protein product fromkaniwa.

BACKGROUND

Bitter taste and low protein levels mark current protein productsderived from quinoa and other pseudocereals. Low water solubility leadsto larger particulates that do not dissolve. The low yield of proteinextract per ounce of raw material results in a higher price per pound ofprotein product. Current extraction methods also degrade the quality ofthe protein, permit unwanted autooxidation, and provide an incompletebalance of essential and non-essential amino acids. What is needed,therefore, is a protein extraction method that provides the desiredprotein levels, amino acid balance, extraction efficiency, andsolubility, without bitterness.

SUMMARY

The present disclosure relates methods for isolating protein fromkaniwa. Particularly, kaniwa does not contain the bitter-tastingcompounds found in quinoa, so starting with kaniwa reduces thepossibility of extracting a bitter-tasting protein product. Furthermore,treating the milled kaniwa with a salt solution allows for the proteinto be precipitated from the protein extract to provide a protein productin better yield with an improved taste and more ready availability infood or commercial applications. This method also provides higherprotein levels, better quality protein levels with an ideal amino acidbalance, and greater water solubility.

In some embodiments, a method for making a protein product from kaniwa.The method comprises (a) milling the kaniwa. (b) The milled kaniwa iscontacted with a salt solution. (c) A protein is precipitated from thekaniwa of step (b). And the precipitated protein is dried to form aprotein product. The protein product may be substantially free ofbitter-tasting compounds, such as saponins. The milling step maycomprise dry milling or wet milling.

The salt may be selected from the Hofmeister series (lyotropic series)based on the ability of its constituent ions to salt out proteins. Inexemplary embodiments, the salt may be selected from the groupconsisting of ammonium chloride, potassium chloride, sodium chloride,calcium chloride, and sodium citrate. In exemplary embodiments, the saltmay be sodium citrate. In particular, the sodium citrate solution ofabout 0.5 wt. % to about 5 wt. % in water and is used in an about 1:5 toabout 1:15 w/v ratio with the milled kaniwa. To effect precipitation ofthe protein, the pH of the salt solution may be adjusted to a pH ofabout 3 to about 4.5.

The contacting step may occur for about 2 to about 6 hours. The dryingstep may occur at about 50° C. to about 60° C. for about 14 to about 18hours. In exemplary embodiments, the moisture content of the driedprotein product may be less that 5 wt. %. In some embodiments, themethod may further comprise centrifugation. In particular embodiments,the protein product may contain a complete set of essential amino acids.

In further embodiments, this disclosure provides a protein productproduced according to the method disclosed herein. A number of consumerproducts may contain the protein product, for example a protein powder,protein drink, protein pill, protein bar, meal replacement, pediatricproduct, or sports nutrition product.

In other embodiments, this disclosure provides a method for making aprotein product from kaniwa. The method comprises (a) contacting kaniwafor 2 to 6 hours with a salt solution of 0.5 wt. % to 5 wt. % in waterand in a 1:5 to 1:15 w/v ratio with the kaniwa. (b) The pH of the saltsolution is adjusted to a pH of 3 to 4.5 to effect precipitation of theprotein from the kaniwa of step (a).

In some other embodiments, this disclosure provides a method for makinga protein product from kaniwa. The method comprises (a) dry milling thekaniwa. (b) The milled kaniwa is contacted for 2 to 6 hours with asodium citrate solution of 0.5 wt. % to 5 wt. % in water and in a 1:5 to1:15 w/v ratio with the milled kaniwa. (c) The pH of the salt solutionis adjusted to a pH of 3 to 4.5 to effect precipitation of the proteinfrom the kaniwa of step (b). (d) The mixture of milled kaniwa and thesalt solution of step (c) is centrifuged to separate the precipitatedprotein from the salt solution. (e) The precipitated protein is dried at50° C. to 60° C. for 14 to 18 hours to form a protein product. Usingthis method, the protein product is substantially free of bitter-tastingcompounds, has a complete set of essential amino acids, and has amoisture content less that 5 wt. %.

Unless otherwise defined, all technical and scientific terms used hereinhave the same meaning as is commonly understood by one of skill in theart to which this disclosure belongs at the time of filing. Ifspecifically defined, then the definition provided herein takesprecedent over any dictionary or extrinsic definition. Further, unlessotherwise required by context, singular terms shall include pluralities,and plural terms shall include the singular. Herein, the use of “or”means “and/or” unless stated otherwise. All patents and publicationsreferred to herein are incorporated by reference.

DETAILED DESCRIPTION

The present disclosure provides a method for disrupting the matrix ofkaniwa, thereby producing a higher protein yield from kaniwa than fromother seeds or grains. To extract, the kaniwa may be dry milled or wetmilled. Optionally, sulfur dioxide may be used during wet milling. Aftermilling, the kaniwa may be treated with a lye or a salt solution. Next,the kaniwa protein may be solvent oil extracted. In other embodiments,protein may be precipitated from the extract by adjusting the pH to a pHof about 3 to about 4.5. Methods to concentrate the precipitatedproteins, include, but are not limited to drying methods (e.g. spraydrying, air drying, drum drying) and centrifugation followed by drying.Protein concentrate and isolate produced according to this disclosurewere analyzed for amino acid composition and functional properties,especially for solubility, water-holding capacity, foaming ability, andtaste. A relatively large quantity of kaniwa may be used in the process.

“Protein product” refers to a protein concentrate or a protein isolate.A protein concentrate is a mixture of protein derived from a feedstock,such as kaniwa, and having a higher concentration per unit volume thanthe feedstock. A protein concentrate may contain other materials, suchas carbohydrates, fats, nucleic acids, vitamins, minerals, and water. Aprotein isolate consists essentially of protein derived from afeedstock. While a protein isolate may contain trace amounts of othernutrients and solvent, it is essentially protein.

“Amino acid” refers to an organic compound having an amino group and acarboxylic acid attached to the same carbon atom. The carbon atom isoptionally substituted with an organic substituent, also referred to asa side-chain. When joined together through amide (peptide) bonds, theseamino acids form proteins. Twenty-three amino acids are proteinogenic,acting as basic building blocks for the proteins. Twenty-one of theseamino acids are “standard” in humans, encoded directly by triplet codonsin the genetic code. Nine of the standard amino acids are “essentialamino acids” in humans, meaning that they cannot be created from othercompounds in the human body. The essential amino acids are histidine,isoleucine, leucine, lysine, methionine, phenylalanine, threonine,tryptophan, and valine. The remaining eleven standard amino acids are“non-essential amino acids,” meaning that they can typically besynthesized from other compounds in the human body. The non-essentialamino acids are alanine, arginine, asparagine, aspartic acid, cysteine,glutamic acid, glutamine, glycine, ornithine, proline, serine, andtyrosine. In an exemplary embodiment, the protein product contains acomplete set of essential amino acids. In other words, the proteinproduct contains some of each of the nine essential amino acids.

“Grain” refers to any cultivated cereal crop used as food. As usedherein, “grain” also encompasses seeds or kernels that may be used orcooked like a grain—so-called “psuedocereals.” “Cereal” refers to agrass (members of the moncot family Poaceae, also known as Gramineae)cultivated for the edible components of their grain composed of theendosperm, germ, and bran. Botanically, these edible components arecalled a caryopsis. In their natural form as a whole grain, they are arich source of vitamins, minerals, carbohydrates, fats, oils, andprotein. Examples of cereals include, but are not limited to, rice,wheat, millet, maize, barley, sorghum, oats, triticale, rye, fonio,spelt, einkorn, emmer, durum, and kamut. Examples of pseudocerealsinclude quinoa, buckwheat, teff, amaranth, chia, and kaniwa. The methodsdescribed herein may be applied to any pseudocereal, particularly tokaniwa.

“Kaniwa” or “kalliwa,” is typically cultivated in the Andes Mountains,and is similar to quinoa with a nutty flavor. But kaniwa is without anybitterness. Though tinier and less fluffy than quinoa, kaniwa has apleasant crunch and makes a satisfying alternative grain. Kaniwa alsohas higher protein content than quinoa, and contains fiber, iron,calcium, and zinc.

Quinoa contains bitter-tasting compounds, such as saponins, that aresubstantially absent from kaniwa. Saponins are often extracted alongwith proteins during processing. Starting from kaniwa instead of quinoaprovides the advantage of no or substantially few bitter-tastingcompounds, thus no or substantially little bitterness. Kaniwa also hasthe advantage of a higher protein content over quinoa, which permitsmore efficient extraction from feedstock.

“Saponin” refers an amphipathic glycoside, which causes soap-likefoaming when shaken in aqueous solutions. Structurally, saponins containa hydrophilic glycoside moiety and a lipophilic triterpene derivative.Typically, saponins are derived from plant material, such as grains, forexample quinoa and amaranth. Saponins are not present in kaniwa.

The protein may be precipitated from a solution. The ionic strength ofthe solution may be controlled, enabling salting out. The pH of thesolution may be controlled, permitting isoelectric precipitation. Thedielectric constant of the solution may be controlled, permittingsolvent fractionation. The temperature of the solution may becontrolled, determining the amount of protein denaturation. The successof a protein precipitation is defined by Leco Nitrogen analysis.

In some embodiments, the milled kaniwa may be contacted with lye. “Lye”refers to any strong alkali, which is highly soluble in water and mayproduce caustic basic solution. In some embodiments, lye may refer tosodium hydroxide, potassium hydroxide, and mixtures thereof. In otherembodiments, the milled kaniwa may be contacted with a salt solution. Inparticular embodiments, the salt may be selected from the groupconsisting of ammonium chloride, potassium chloride, sodium chloride,calcium chloride, and sodium citrate. In exemplary embodiments, the saltmay be sodium citrate. The salt solution may have a concentrationranging from about 0.5 wt. % to about 5 wt. % in water, such as fromabout 0.5 wt. % to about 1 wt. %, from about 1 wt. % to about 1.5 wt. %in water, from about 1.5 wt. % to about 2 wt. % in water, from about 2wt. % to about 2.5 wt. % in water, from about 2.5 wt. % to about 3 wt. %in water, from about 3 wt. % to about 3.5 wt. % in water, from about 3.5wt. % to about 4 wt. % in water, from about 4 wt. % to about 4.5 wt. %in water, from about 4.5 wt. % to about 5 wt. % in water. In exemplaryembodiments, the salt solution may have a contraction of 1 wt. % or 3wt. % in water.

The ratio of salt solution to milled kaniwa can and will vary. In someembodiments, the salt solution may be used in an about 1:5 to about 1:15w/v ratio with the milled kaniwa, for example in an about 1:5 to about1:6 w/v ratio, in an about 1:6 to about 1:7 w/v ratio, in an about 1:7to about 1:8 w/v ratio, in an about 1:8 to about 1:9 w/v ratio, in anabout 1:9 to about 1:10 w/v ratio, in an about 1:10 to about 1:11 w/vratio, in an about 1:11 to about 1:12 w/v ratio, in an about 1:12 toabout 1:13 w/v ratio, in an about 1:13 to about 1:14 w/v ratio, or in anabout 1:14 to about 1:15 w/v ratio. In exemplary embodiments, the saltsolution may be used in an about 1:10 w/v ratio with the milled kaniwa.

The contacting step between the milled kaniwa and the salt solution mayoccur for about 2 hours to about 6 hours, such as about 2 hours, about 3hours, about 4 hours, about 5 hours, or about 6 hours. The contact stepmay comprise skaking, stifling, or agitating the mixture of milledkaniwa and salt solution.

To effect precipitation of the protein, the pH of the salt solution maybe adjusted to a pH of 3 to 4.5, for example to a pH of about 3, a pH ofabout 3.5, a pH of about 4, or a pH of about 4.5. In exemplaryembodiments, the salt is sodium citrate with a concentration of about 1wt. % and a pH of about 4. In other exemplary embodiments, the salt issodium citrate with a concentration of about 3 wt. % and a pH of about3.5. The pH may be adjusted using a food quality proton donor oracceptor, such as hydrochloric acid or sodium hydroxide, using anymethod known in the art.

In other embodiments, the protein product may be extracted from solutionusing a suitable organic solvent. Non-limiting examples of suitableaprotic solvents include acetone, acetonitrile, diethoxymethane,N,N-dimethylformamide (DMF), dimethyl sulfoxide (DMSO),N,N-dimethylpropionamide, 1,3 -dimethyl-3 ,4,5 ,6-tetrahydro-2(1H)-pyrimidinone (DMPU), 1,3-dimethyl-2-imidazolidinone (DMI),1,2-dimethoxyethane (DME), dimethoxymethane, bis(2-methoxyethyl)ether,N,N-dimethylacetamide (DMAC), 1,4-dioxane, N-methyl-2-pyrrolidinone(NMP), ethyl acetate, ethyl formate, ethyl methyl ketone, formamide,hexachloroacetone, hexamethylphosphoramide, methyl acetate,N-methylacetamide, N-methylformamide, methylene chloride, nitrobenzene,nitromethane, propionitrile, sulfolane, tetramethylurea, tetrahydrofuran(THF), 2-methyl tetrahydrofuran, trichloromethane, and combinationsthereof. Suitable examples of protic solvents include, but are notlimited to, methanol, ethanol, isopropanol, n-propanol, isobutanol,n-butanol, s-butanol, t-butanol, formic acid, acetic acid, water, andcombinations thereof. Suitable organic solvents include, but are notlimited to, alkane and substituted alkane solvents (includingcycloalkanes), aromatic hydrocarbons, esters, ethers, ketones,combinations thereof, and the like. Specific organic solvents that maybe employed, include, for example, acetonitrile, benzene, butyl acetate,t-butyl methylether, t-butyl methylketone, chlorobenzene, chloroform,chloromethane, cyclohexane, dichloromethane, dichloroethane, diethylether, ethyl acetate, diethylene glycol, fluorobenzene, heptane, hexane,isobutylmethylketone, isopropyl acetate, methylethylketone,methyltetrahydrofuran, pentyl acetate, n-propyl acetate,tetrahydrofuran, toluene, and combinations thereof. In exemplaryembodiments, the solvent used for protein extraction may be diethylether or ethanol.

In some embodiments, the method may further comprise centrifugation toseparate the precipitated protein from the supernatant. Generally, whenused, centrifugation would occur before the drying step. Thecentrifugation speed may range from about 7,000 rpm to about 28,000 rpm,such as from about 7,000 rpm to about 14,000 rpm, from about 14,000 rpmto about 21,000 rpm, or from about 21,000 rpm to about 28,000 rpm. Thecentrifugation time may range from about 5 minutes to about 5 hours,such as from about 5 minutes to about 15 minutes, from about 15 minutesto about 30 minutes, from about 30 minutes to about 1 hour, from about 1hour to about 2 hours, from about 2 hours to about 3 hours, from about 3hours to about 4 hours, or from about 4 hours to about 5 hours. Thecentrifugation temperature may range from about 0° C. to about 15° C.,such as from about 0° C. to about 5° C., from about 2° C. to about 5°C., from about 5° C. to about 10° C., or from about 10° C. to about 15°C. In exemplary embodiments, the centrifugation may occur at about14.000 rpm for about 30 min at about 4° C.

The drying step may use any method known in the art. The dryingtemperature may range from about 25° C. to about 100° C., such as fromabout 25° C. to about 30° C., such as from about 30° C. to about 35° C.,from about 35° C. to about 40° C., from about 40° C. to about 45° C.,from about 45° C. to about 50° C., from about 50° C. to about 55° C.,from about 55° C. to about 60° C., from about 60° C. to about 65° C.,from about 65° C. to about 70° C., from about 70° C. to about 75° C.,from about 75° C. to about 80° C., from about 80° C. to about 85° C.,from about 85° C. to about 90° C., from about 90° C. to about 95° C., orfrom about 95° C. to about 100° C. In exemplary embodiments, the dryingtemperature may be from about 50° C. to about 60° C., such as about 55°C.

The drying step may occur for about 12 hours to about 24 hours, such asfor about 12 hours to about 14 hours, for about 14 hours to about 16hours, for about 16 hours to about 18 hours, for about 18 hours to about20 hours, for about 20 hours to about 22 hours, or for about 22 hours toabout 24 hours. In exemplary embodiments, the drying step may occur forabout 14 to about 18 hours, such as about 16 hours. In exemplaryembodiments, the moisture content of the dried protein product may beless that about 10 wt. %, such as less than about 5 wt. %, less thanabout 4 wt. %, less than about 3 wt. %, less than about 2 wt. %, lessthan about 1 wt. %, less than about 0.5 wt. %, less than about 0.1 wt.%, or effectively 0 wt. % (that is, trace amounts of water or below thelimit of detection).

A protein product as described herein may be used in any common proteinsupplement. Protein supplements come in various forms, including, forexample, protein powders, protein drinks, protein pills, protein bars,meal replacements, pediatric products, and sports nutrition products.For example, a kaniwa protein product providing a complete set of aminoacids may be substituted for soy protein as the ingredient in anyprotein supplement. In some embodiments, the protein supplement (e.g., apowder, pill, bar, and the like, as described herein) may comprise about5-40 grams of kaniwa protein product per serving (e.g., equal to,greater than, at least, or any number in between 5, 10, 15, 20, 25, 30,35, 40, 45, or 50 grams of protein/serving) wherein a serving of thecomposition may be less than, greater than, at least, or any number inbetween 10, 10.5, 11, 11.5, 12, 12.5, 13, 13.5, 14, 14.5, 15, 15.5, 16,16.5, 17, 17.5, 18, 18.5, 19, 19.5, 20, 30, 40, 50, 60, 70, 80, 90 or100 grams.

The kaniwa protein product may be used in a protein powder. The proteinpowder may further comprise one or more excipients, such as fillers,solublizers, flavorants, sweeteners, dietary minerals, binders,vitamins, fatty acids (e.g., omega-3 or omega-6 fatty acids), caffeine,and the like. The protein powders may be formulated to provide a proteindrink upon the addition of water and adequate mixing, such as vigorousshaking.

In other embodiments, the protein product may be formulated as a proteindrink in stable liquid form. The protein drink may comprise more thanabout 10 grams of kaniwa protein per 100 mL. The protein drink may havea pH of from about 5.0 to about 8.0, such as about 7.0, or about 6.8.The protein drink may further comprise one or more excipients, such aswater, fillers, solublizers, flavorants, sweeteners, dietary minerals,binders, vitamins, fatty acids (e.g., omega-3 or omega-6 fatty acids),caffeine and the like.

In other embodiments, the protein product may be formulated as a proteinbar, chew, snack, or other food item. The protein bar may furthercomprise one or more excipients, such as fillers, solublizers,flavorants, sweeteners, dietary minerals, binders, vitamins, fatty acids(e.g., omega-3 or omega-6 fatty acids), caffeine, and the like. Theprotein bar may also comprise nuts, dried fruit, pretzels, soybeans,chocolate, coffee, coconut, and other ingredients commonly found inprotein bars.

In particular embodiments, the protein powder, protein drink, proteinbar may be formulated for pediatric use, such that it may be used abreastmilk substitute, infant formula, infant formula component, orpediatric food fortifier. In still other embodiments, the proteinpowder, protein drink, or protein bar may be formulated as a sportsnutrition product which provides to the person the amount of proteinused for a certain activity level. For example, recreational athletesuse about 0.5 to about 0.75 grams of protein daily for every per poundof bodyweight, competitive athletes use about 0.6 to about 0.9 grams perpound of bodyweight, teenage athletes use about 0.8 to about 0.9 gramsper pound of bodyweight, and athletes building muscle mass use about 0.7to about 0.9 grams per pound of bodyweight. In some embodiments, theprotein powder, protein drink, or protein bar may be used as a mealreplacement.

In still other embodiments, the protein product may be formulated as aprotein pill. The protein pill may further one or more excipients, suchas comprise fillers, solublizers, flavorants, sweeteners, dietaryminerals, binders, vitamins, fatty acids (e.g., omega-3 or omega-6 fattyacids), caffeine, and the like.

In one embodiment, the excipient may be a binder, which holds theprotein supplement together until administration. Suitable bindersinclude starches, pregelatinized starches, gelatin,polyvinylpyrrolidone, cellulose, methylcellulose, sodiumcarboxymethylcellulose, ethylcellulose, polyacrylamides,polyvinyloxoazolidone, polyvinyl alcohols, C₁₂-C₁₈ fatty acid alcohol,polyethylene glycol, polyols, saccharides, oligosaccharides,polypeptides, peptides, and combinations thereof.

The excipient may include a flavorant that may act as a flavoring agentand/or as a flavor-masking agent. In some embodiments, the flavorant maycomprise one or more of a sweetening agent, a savory agent (i.e., anagent that imbues the protein supplement with a salty flavor), abittering agent, and a souring agent. Flavorants may be chosen fromsynthetic flavor oils and flavoring aromatics and/or natural oils,extracts from plants, leaves, flowers, fruits, and combinations thereof.By way of example, these may include cinnamon oils, oil of wintergreen,peppermint oils, clover oil, hay oil, anise oil, eucalyptus, vanilla,citrus oils (such as lemon oil, orange oil, grape and grapefruit oil),and fruit essences (such as apple, peach, pear, strawberry, raspberry,cherry, plum, pineapple, and apricot). In some embodiments, theflavoring agents and/or flavor-masking agents may comprise avanilla-comprising composition, for example ethylvanillin, vanillin-RHD,vanillin-Merck, vanilla-TG-old; chocolate, coffee, and suitable solvents(e.g., ethanol and/or water).

In another embodiment, the excipient may include a sweetener. By way ofnon-limiting example, the sweetener may be selected from glucose (cornsyrup), dextrose, invert sugar, fructose, and mixtures thereof (when notused as a carrier); saccharin and its various salts such as the sodiumsalt; dipeptide sweeteners such as aspartame; dihydrochalcone compounds,glycyrrhizin; stevia-derived sweeteners; chloro derivatives of sucrosesuch as sucralose; sugar alcohols such as sorbitol, mannitol, xylitol,and the like. Also contemplated are hydrogenated starch hydrolysates andthe synthetic sweetener3,6-dihydro-6-methyl-1,2,3-oxathiazin-4-one-2,2-dioxide, particularlythe potassium salt (acesulfame-K), and sodium and calcium salts thereof.In particular embodiments, the protein supplement may be formulated fororal administration and include one or more of the following flavorant(e.g., sweetening agents): sucralose, MagnaSweet®, Di-Pac® compressiblesugar (i.e., a 97:3 mixture of sucrose and maltodextrin), ThaumatinT200X, Talin-Pure, OptisweetSD, stevia extract rebaudioside A, and/orneotame.

The excipient may include a taste-masking agent. Taste-masking materialsinclude cellulose hydroxypropyl ethers (HPC); low-substituted cellulosehydroxypropyl ethers (L-HPC); cellulose hydroxypropyl methyl ethers(HPMC); methylcellulose polymers and mixtures thereof; polyvinyl alcohol(PVA); hydroxyethylcelluloses; carboxymethylcelluloses and saltsthereof; polyvinyl alcohol and polyethylene glycol co-polymers;monoglycerides or triglycerides; polyethylene glycols; acrylic polymers;mixtures of acrylic polymers with cellulose ethers; cellulose acetatephthalate; and combinations thereof.

In some embodiments, the flavorant may comprise a percent weight perfinal volume of the protein supplement form about 50% to about 0.001%,depending on the agent selected, such as from about 40% to about 0.01%,from about 30% to about 0.01%, from about 1% to about 30%, or from about5% to about 15%. As previously mentioned, the protein supplement mayinclude more than one flavorant.

In another embodiment, the excipient may be a filler, which adds bulk tothe protein supplement for easier handling and more accurate dosing.Suitable fillers include carbohydrates, inorganic compounds, andpolyvinylpyrrolidone. By way of non-limiting example, the filler may becalcium sulfate, both di- and tri-basic, starch, calcium carbonate,magnesium carbonate, microcrystalline cellulose, dibasic calciumphosphate, magnesium carbonate, magnesium oxide, calcium silicate, talc,modified starches, lactose, sucrose, mannitol, and sorbitol.

The excipient may be a non-effervescent disintegrant, which allows theprotein supplement to more easily dissolve after administration withoutevolving gas. Suitable examples of non-effervescent disintegrantsinclude starches (such as corn starch, potato starch, and the like),pregelatinized and modified starches thereof, sweeteners, clays (such asbentonite), microcrystalline cellulose, alginates, sodium starchglycolate, and gums (such as agar, guar, locust bean, karaya, pecitin,and tragacanth).

The excipient may comprise a preservative, which increases the stabilityand storage lifetime of the protein supplement, particularly delayingunwanted degradation of the active ingredients. Suitable examples ofpreservatives include antioxidants (such as alpha-tocopherol orascorbate) and antimicrobials (such as parabens, chlorobutanol orphenol). In other embodiments, an antioxidant such as butylatedhydroxytoluene (BHT) or butylated hydroxyanisole (BHA) may be used.

The excipient may include a diluent, which diminishes the relativeconcentrations of other components within the protein supplement.Diluents suitable for use include food-grade saccharides such assucrose, dextrose, lactose, microcrystalline cellulose, fructose,xylitol, and sorbitol; polyhydric alcohols; starches; pre-manufactureddirect compression diluents; and mixtures of any of the foregoing.

The excipient may comprise a surfactant, which alters the solubilityparameters of the other components within the protein supplement. Invarious embodiments, the surfactant may be a alkylaryl polyetheralcohol, such as Triton™ X-100, Surfonic™ N-100 (nonoxaynol-10), orWitconol™ NP-100; or a poloxamer, such as Pluronic™, Synperonic™, orKolliphor™. Other suitable examples of surfactants include, for example,2-acrylamido-2-methylpropane sulfonic acid, alkyl polyglycoside,ammonium perfluorononanoate, benzalkonium chloride (BAC), benzethoniumchloride (BZT), 5-bromo-5-nitro-1,3-dioxane, cetyl trimethylammoniumbromide (CTAB, hexadecyltrimehtylammonium bromide, cetyltrimethylammonium chloride), cetylpridinium chloride (CPC),cyclohexyl-1-hexyl-maltopyranoside, decylmaltopyranoside, decylpolyglucose, dimethyldioctadecylammonium chloride,dioctadecyldimethylammmonium bromide (DODAB),dipalmitoylphosphatidylcholine, lauryldimethylamine oxide,dodecylmaltopyranoside, magnesium laureth sulfate polyethoxylated tallowamine (POEA), octenidine dihydrochloride, octylphenoxypolyethoxyethanol(Igepal™ CA-630), octylthioglucopyranoside (OTG), ox gall, sodiumnonanoyloxybenzensulfonate, sorbitan monolaurate, surfactin, andthonozonium bromide. In exemplary embodiments, the surfactant may be apoloxamer or sodium lauryl sulfate.

The excipient may be a lubricant, which allows easier removal of theprotein supplement from molds during manufacture and may aidadministration of the protein supplement. Suitable non-limiting examplesof lubricants include magnesium stearate, calcium stearate, zincstearate, hydrogenated vegetable oils, sterotex, polyoxyethylenemonostearate, talc, polyethyleneglycol, sodium benzoate, sodium laurylsulfate, magnesium lauryl sulfate, and light mineral oil.

The excipient may be a dispersion enhancer, which aids dispersion of thecomponents of the protein supplement within the subject afteradministration. Suitable dispersants may include starch, alginic acid,polyvinylpyrrolidones, guar gum, kaolin, bentonite, purified woodcellulose, sodium starch glycolate, isoamorphous silicate, andmicrocrystalline cellulose.

Depending upon the embodiment, it may be desirable to provide a coloringagent, which aids cosmetic appeal, visualization, and identification ofthe protein supplment. Suitable color additives include food, drug andcosmetic colors (FD&C), drug and cosmetic colors (D&C), or external drugand cosmetic colors (Ext. D&C). These colors or dyes, along with theircorresponding lakes, and certain natural and derived colorants may besuitable for use in the present invention depending on the embodiment.

In various embodiments, the excipient may include a pH modifier, whichmay alter the solubility profile and bioavailability parameters ofcomponents within the protein supplement. In certain embodiments, the pHmodifier may include sodium carbonate or sodium bicarbonate.

The weight fraction of the excipient or combination of excipients in theprotein supplement may be about 98% or less, about 95% or less, about90% or less, about 85% or less, about 80% or less, about 75% or less,about 70% or less, about 65% or less, about 60% or less, about 55% orless, about 50% or less, about 45% or less, about 40% or less, about 35%or less, about 30% or less, about 25% or less, about 20% or less, about15% or less, about 10% or less, about 5% or less, about 2%, or about 1%or less of the total weight of the protein supplement.

As used herein, the terms “about” and “approximately” designate that avalue is within a statistically meaningful range. Such a range can betypically within 20%, more typically still within 10%, and even moretypically within 5% of a given value or range. The allowable variationencompassed by the terms “about” and “approximately” depends on theparticular system under study and can be readily appreciated by one ofordinary skill in the art.

As used herein, the term “w/w” designates the phrase “by weight” and isused to describe the concentration of a particular substance in amixture or solution.

Although the disclosure described herein is susceptible to variousmodifications and alternative iterations, specific embodiments thereofhave been described in greater detail above. It should be understood,however, that the detailed description is not intended to limit thedisclosure to the specific embodiments disclosed. Rather, it should beunderstood that the disclosure is intended to cover all modifications,equivalents, and alternatives falling within the spirit and scope of thedisclosure as defined by the claim language.

EXAMPLES

The following examples are included to demonstrate certain embodimentsof the disclosure. It should be appreciated by those of skill in the artthat the techniques disclosed in the examples represent techniquesdiscovered by the inventors to function well in the practice of thedisclosure. Those of skill in the art should, however, in light of thepresent disclosure, appreciate that many changes can be made in thespecific embodiments that are disclosed and still obtain a like orsimilar result without departing from the spirit and scope of thedisclosure, therefore all matter set forth is to be interpreted asillustrative and not in a limiting sense.

Example 1—Extracting Protein from Kaniwa

Kaniwa was milled in a variety of mills available for small scaleprocesses: hammer min, coffee grinder, and kitchen counter top grainmill. The ionic strength, the pH of the solution, dielectric constant,and temperature of the solution were controlled to affect the proteinprecipitation. In particular, the milled kaniwa was treated with a 1% ora 3% aqueous sodium citrate solution at a 1:10 w/v ratio. Next, themixture was shaken on a laboratory shaker for 2 to 6 hours, Aftershaking, the citrate solution with the extract was adjusted to a pH of3.5 to 4 to precipitate the protein. in particular, a 1% aqueous sodiumcitrate solution was used with a pH of 4.0 to effect precipitation ofthe protein product from the citrate extract, or a 3% aqueous sodiumcitrate solution was used with a pH 3.5 to effect precipitation. Theprotein precipitation was measured by Leco Nitrogen analysis.

In either case, after adequate time had passed, the citrate extract wascentrifuged at 14,000 rpm for 30 min at 4° C. to obtain a pellet of theprecipitated protein. The precipitated protein was dried in a forced aircabinet at 55° C. for 16 hours, thereby providing a final moisturecontent of less than 5 wt. %. The dried protein product was analyzed foramino acid composition and functional properties, especially forsolubility, water-holding capacity, foaming ability, and taste. Theprotein product was low in bitterness as demonstrated by taste testing.The protein product also had a complete set of essential amino acids,and had good water solubility.

Example 2—Pretreatment with of Kaniwa with Lye

Pellets of kaniwa were soaked in a 0.1 N sodium hydroxide solution forvarious periods of time. The product was rinsed several times withpotable water, dried, milled and the protein extracted as describedabove in Example 1. This procedure provided a pH adjustment to theprotein product extracted from the kaniwa.

Example 3—Wet milling kaniwa to extract protein

A portion of kaniwa was wet milled. Optionally, sulfur dioxide was usedduring milling. After milling, the kaniwa was treated with lye solution,comprising water and sodium hydroxide. Next, the kaniwa was solvent oilextracted. Two solvents were tested for defatting kaniwa: diethyl etherand ethanol. Each solvent was equally efficient. Use of ethanol forextraction increased the protein yield when other salts were used. Withsodium citirate, however, ethanol slightly decreased the protein yield.The extraction was performed on an about 1:10 w/v ration at roomtemperature using a laboratory shaker for 2 to 6 hours.

The protein was precipitated from a solvent oil solution. The proteinprecipitation was measured by Leco Nitrogen analysis. The precipitatedproteins were further concentrated by drying an analyzed as describedabove in Example 1. Lye, sulfites, and sodium bicarbonate were only usedfor the wet milling process. None of them was successful in increasingthe yield of the protein extracted as compared to extraction methodsknown in the art.

While specific embodiments have been described above with reference tothe disclosed embodiments and examples, such embodiments are onlyillustrative and do not limit the scope of the disclosure. Changes andmodifications can be made in accordance with ordinary skill in the artwithout departing from the disclosure in its broader aspects as definedin the following claims.

What is claimed is:
 1. A method for making a protein product fromkaniwa, comprising: (a) milling the kaniwa; (b) contacting the milledkaniwa with a salt solution; (c) precipitating a protein from the kaniwaof step (b); and (d) drying the precipitated protein to form a proteinproduct.
 2. The method of claim 1, wherein the protein product issubstantially free of bitter-tasting compounds.
 3. The method of claim2, wherein the bitter-tasting compound is a saponin.
 4. The method ofclaim 1, wherein the milling step comprises at least one selected fromthe group consisting of dry milling and wet milling.
 5. The method ofclaim 1, wherein the salt is selected from the group consisting ofammonium chloride, potassium chloride, sodium chloride, calciumchloride, and sodium citrate.
 6. The method of claim 5, wherein the saltis sodium citrate.
 7. The method of claim 6, wherein the sodium citratesolution of 0.5 wt. % to 5 wt. % in water and is used in a 1:5 to 1:15w/v ratio with the milled kaniwa.
 8. The method of claim 1, furthercomprising adjusting the pH of the salt solution to a pH of 3 to 4.5 toeffect precipitation of the protein.
 9. The method of claim 6, whereinthe contacting step occurs for 2 to 6 hours.
 10. The method of claim 1,wherein the drying step occur at 50° C. to 60° C. for 14 to 18 hours.11. The method of claim 10, wherein the moisture content of the driedprotein product is less that 5 wt. %.
 12. The method of claim 1, furthercomprising centrifugation.
 13. The method of claim 1, further comprisingcontaining the kaniwa with a solution of lye before milling.
 14. Themethod of claim 1, wherein the protein product contains a complete setof essential amino acids.
 15. A protein product produced according tothe method of claim
 1. 16. A protein powder containing a protein productproduced according to the method of claim
 1. 17. A protein drinkcontaining protein product produced according to the method of claim 1.18. A protein pill containing a protein product produced according tothe method of claim
 1. 19. A protein bar containing a protein productproduced according to the method of claim
 1. 20. A meal replacementcontaining a protein product produced according to the method ofclaim
 1. 21. A pediatric product containing a protein product producedaccording to the method of claim
 1. 22. A sports nutrition productcontaining a protein product produced according to the method ofclaim
 1. 23. A method for making a protein product from kaniwa,comprising: (a) contacting kaniwa for 2 to 6 hours with a salt solutionof 0.5 wt. % to 5 wt. % in water and in a 1:5 to 1:15 w/v ratio with thekaniwa; (b) adjusting the pH of the salt solution to a pH of 3 to 4.5 toeffect precipitation of the protein from the kaniwa of step (a).
 24. Themethod of claim 23, wherein the protein product is substantially free ofbitter-tasting compounds.
 25. The method of claim 23, wherein thebitter-tasting compound is a saponin.
 26. The method of claim 23,further comprising milling the kaniwa before step (a), wherein themilling step comprises at least one selected from the group consistingof dry milling and wet milling.
 27. The method of claim 23, wherein thesalt is selected from the group consisting of ammonium chloride,potassium chloride, sodium chloride, calcium chloride, and sodiumcitrate.
 28. The method of claim 27, wherein the salt is sodium citrate.29. The method of claim 23, further comprising drying the precipitatedprotiein at 50° C. to 60° C. for 14 to 18 hours.
 30. The method of claim29, wherein the moisture content of the dried protein product is lessthat 5 wt. %.
 31. The method of claim 23, further comprisingcentrifugation.
 32. The method of claim 23, further comprisingcontaining the kaniwa with a solution of lye before milling.
 33. Themethod of claim 23, wherein the protein product contains a complete setof essential amino acids.
 34. A method for making a protein product fromkaniwa, comprising: (a) dry milling the kaniwa; (b) contacting themilled kaniwa for 2 to 6 hours with a sodium citrate solution of 0.5 wt.% to 5 wt. % in water and in a 1:5 to 1:15 w/v ratio with the milledkaniwa; (c) adjusting the pH of the salt solution to a pH of 3 to 4.5 toeffect precipitation of the protein from the kaniwa of step (b); (d)centrifuging the mixture of milled kaniwa and the salt solution of step(c) to sepeate the precipitated protein from the salt solution; and (e)drying the precipitated protein at 50° C. to 60° C. for 14 to 18 hoursto form a protein product; wherein the protein product is substantiallyfree of bitter-tasting compounds, has a complete set of essential aminoacids, and has a moisture content less that 5 wt. %.